KIFS as Modifiers of the RHO Pathway and Methods of Use

Abstract

Human KIF23 genes are identified as modulators of the RHO pathway, and thus are therapeutic targets for disorders associated with defective RHO function. Methods for identifying modulators of RHO, comprising screening for agents that modulate the activity of KIF23 are provided.

1 . A method of identifying a candidate RHO pathway modulating agent, said method comprising the steps of:
 (a) providing an assay system comprising an KIF23 polypeptide or nucleic acid;   (b) contacting the assay system with a test agent under conditions whereby, but for the presence of the test agent, the system provides a reference activity; and   (c) detecting a test agent-biased activity of the assay system, wherein a difference between the test agent-biased activity and the reference activity identifies the test agent as a candidate RI-10 pathway modulating agent.   
     
     
         2 . The method of  claim 1  wherein the assay system comprises cultured cells that express the KIF23 polypeptide. 
     
     
         3 . The method of  claim 2  wherein the cultured cells additionally have defective RHO function. 
     
     
         4 . The method of  claim 1  wherein the assay system includes a screening assay comprising a KIF23 polypeptide, and the candidate test agent is a small molecule modulator. 
     
     
         5 . The method of  claim 4  wherein the assay is a binding assay. 
     
     
         6 . The method of  claim 1  wherein the assay system is selected from the group consisting of an apoptosis assay system, a cell proliferation assay system, and an angiogenesis assay system. 
     
     
         7 . The method of  claim 1  wherein the assay system includes a binding assay comprising a KIF23 polypeptide and the candidate test agent is an antibody. 
     
     
         8 . The method of  claim 1  wherein the assay system includes an expression assay comprising a KIF23 nucleic acid and the candidate test agent is a nucleic acid modulator. 
     
     
         9 . The method of  claim 8  wherein the nucleic acid modulator is an antisense oligomer. 
     
     
         10 . The method of  claim 8  wherein the nucleic acid modulator is a PMO. 
     
     
         11 . The method of  claim 1  additionally comprising:
 (d) administering the candidate RHO pathway modulating agent identified in (c) to a model system comprising cells defective in RHO function and, detecting a phenotypic change in the model system that indicates that the RHO function is restored. 
 
     
     
         12 . The method of  claim 11  wherein the model system is a mouse model with defective RHO function. 
     
     
         13 . A method for modulating a RHO pathway of a cell comprising contacting a cell defective in RHO function with a candidate modulator that specifically binds to a KIF23 polypeptide, whereby RHO function is restored. 
     
     
         14 . The method of  claim 13  wherein the candidate modulator is administered to a vertebrate animal predetermined to have a disease or disorder resulting from a defect in RHO function. 
     
     
         15 . The method of  claim 13  wherein the candidate modulator is selected from the group consisting of an antibody and a small molecule. 
     
     
         16 . The method of  claim 1 , comprising the additional steps of
 (d) providing a secondary assay system comprising cultured cells or a non-human animal expressing KIF23,   (e) contacting the secondary assay system with the test agent of (b) or an agent derived therefrom under conditions whereby, but for the presence of the test agent or agent derived therefrom, the system provides a reference activity; and   (f) detecting an agent-biased activity of the second assay system,   wherein a difference between the agent-biased activity and the reference activity of the second assay system confirms the test agent or agent derived therefrom as a candidate RHO pathway modulating agent,   and wherein the second assay detects an agent-biased change in the RHO pathway.   
     
     
         17 . The method of  claim 16  wherein the secondary assay system comprises cultured cells. 
     
     
         18 . The method of  claim 16  wherein the secondary assay system comprises a non-human animal. 
     
     
         19 . The method of  claim 18  wherein the non-human animal mis-expresses a RHO pathway gene. 
     
     
         20 . A method of modulating RHO pathway in a mammalian cell comprising contacting the cell with an agent that specifically binds a KIF23 polypeptide or nucleic acid. 
     
     
         21 . The method of  claim 20  wherein the agent is administered to a mammalian animal predetermined to have a pathology associated with the RHO pathway. 
     
     
         22 . The method of  claim 20  wherein the agent is a small molecule modulator, a nucleic acid modulator, or an antibody. 
     
     
         23 . A method for diagnosing a disease in a patient comprising:
 obtaining a biological sample from the patient;   contacting the sample with a probe for KIF23 expression;   comparing results from step (b) with a control;   determining whether step (c) indicates a likelihood of disease.   
     
     
         24 . The method of  claim 23  wherein said disease is cancer. 
     
     
         25 . The method according to  claim 24 , wherein said cancer is a cancer as shown in Table 2 as having >25% expression level.